tbuckley/cabot-search huggingface dataset:
Abstract 4210: Gene therapy approach to suppress metastasis in a mouse melanoma model
Authors: Richard Heller, Megan Scott, Cathryn Mangiamele, Jody Synowiec, Guilan Shi
Journal: Cancer Research (2022)

Abstract Immune checkpoint inhibitors (ICIs) have been demonstrated to have significant clinical benefits. Effectiveness of ICIs have been limited by both primary and acquired resistance. In addition, absence of T cell infiltration within the tumor site is one of the major obstacles limiting ICIs efficacy against solid tumors. Interleukin-12 (IL-12) is a potent immune stimulator that has been shown to stimulate growth and survival of T cells as well as NK cells. Utilizing a B16.F10 melanoma model in C57Bl/6 mice, we observed a significant reduction in T regulatory cells and myeloid derived suppressor cells while T effector cells were significantly increased within the tumor microenvironment (TME) following intratumor delivery of a plasmid encoding IL-12 using gene electrotransfer. Monotherapy with plasmid IL-12 delivered with gene electrotransfer (pIL-12 GET) resulted in prolonged disease-free survival as well as long term immune memory. To determine the robustness of this therapeutic approach and to evaluate the responsiveness in combination with anti-PD1, we used a two-tumor model consisting of a subcutaneous B16F10 tumor and B16F10 cells expressing luciferase injected via the intraperitoneal route in a C57Bl/6 mouse. Intratumor delivery of pIL-12 GET as a monotherapy resulted in reduction or elimination of the subcutaneous tumor. However, the monotherapy approach was only successful in reducing the peritoneal spread in about 50% of the mice. When pIL-12 GET was combined with anti-PD1 administered via an intraperitoneal injection not only was there an elimination of the subcutaneous tumor, but it resulted in the elimination of intraperitoneal metastatic growth. The elimination of peritoneal spread was confirmed utilizing an In Vivo Imaging System. Observations on day 60 revealed background levels of luminescence in 90% of mice treated with the combination therapy thus confirming long-term disease-free survival of these mice. This level of response was not seen in mice treated with anti-PD-1 alone or with simple injection of pIL-12. In addition, the combination therapy resulted in an enhanced memory response which protected against growth of new tumors following challenge. In addition, exposure of tumor cells to IL-12 resulted in upregulation of MHC class 1 and PDL1 expression. The results have this study suggest that the combination of pIL-12 GET combined with ICIs could be utilized as an effective combination therapy for melanoma. Citation Format: Richard Heller, Megan Scott, Cathryn Mangiamele, Jody Synowiec, Guilan Shi. Gene therapy approach to suppress metastasis in a mouse melanoma model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 4210.
Abstract 359: Pharmacologically induced polyploidy triggers BCL2 dependence in lymphomas
Authors: Ana Portelinha, Mariana da Silva Ferreira, Tatiana Erazo, Zahra Asgari, Elisa de Stanchina, Anas Younes, Hans-Guido Wendel
Journal: Cancer Research (2022)

Abstract Spontaneous whole genome duplication (WGD) and the adaptive mutations that disrupt cell death and proliferation checkpoints are infrequent events in aggressive B cell lymphomas. This indicates that lymphomas may be especially sensitive to therapeutics that trigger genomic instability. Here, we report the powerful therapeutic combination of Polo-like kinase 4 (PLK4) and BCL2 inhibitors for aggressive lymphomas. Mechanistically, the PLK4 inhibitor CFI-400945 impairs centrosome duplication leading to genomic instability and tetraploidy (4n) in lymphoma cells. Elimination of polyploid lymphoma cells is mediated by the pro-apoptotic BAX protein resulting in increased BCL2 dependence for cell survival. The selective BCL2 inhibitor, venetoclax, acts in a synthetic lethal manner with PLK4 inhibition in both CFI-400945 sensitive and in CFI-400945 resistant lymphomas. As PLK4 is dispensable in non-proliferating cells, synthetic lethality is preferentially observed in tumor cells, while sparing vital organs. Hence, B cell lymphomas are ill-prepared for the rapid, pharmacologic induction of polyploidy resulting in a synthetic and tumor-specific dependency on BCL2. Citation Format: Ana Portelinha, Mariana da Silva Ferreira, Tatiana Erazo, Zahra Asgari, Elisa de Stanchina, Anas Younes, Hans-Guido Wendel. Pharmacologically induced polyploidy triggers BCL2 dependence in lymphomas [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 359.
Abstract 5214: Copper chelation overcomes the immunosuppressive tumor microenvironment in neuroblastoma
Authors: Jourdin R. Rouaen, Daniele Mercatelli, Federica Saletta, Ensieh M. Poursani, Jayne E. Murray, Nicodemus Tedla, Federico M. Giorgi, Orazio Vittorio
Journal: Cancer Research (2022)

Abstract Immunotherapy remains a promising approach in the treatment of high-risk neuroblastoma, however efficacy is hampered by the immunosuppressive tumor microenvironment. We previously reported a link between intratumoral copper levels and Programmed Death-Ligand 1 (PD-L1) expression and therefore sought to elucidate the wider impact of copper chelation on immune evasion strategies (Voli et al 2020). Here we profile key elements of the neuroblastoma tumor microenvironment which work in concert to promote immunosuppression and tumor progression. Using the murine TH-MYCN model, animals were treated daily with the copper chelation agent tetraethylenepentamine (TEPA) for seven days before tumor resection and processing. Transcriptome analyses using single-cell RNA-sequencing revealed copper chelation positively skewed the functional status of tumor, immune and stromal compartments to enhance the anti-tumor immune response. This was supported by a tissue microarray screen using NanoString Digital Spatial Profiling which identified increased immune cell infiltration and regions of active tumor killing. High-resolution mass-spectrometry detected upregulation of major histocompatibility complex (MHC) proteins and exclusive antigenic signatures in peptide repertoires. A multiplex cytokine array indicated an improved anti-tumorigenic response underscored by T cell and Natural Killer cell activation. Importantly, combination of copper chelation and anti-GD2 antibody therapy led to tumor regression and significantly improved survival in vivo. Collectively, this study demonstrates the ability of copper chelation to successfully mitigate tumoral immune evasion strategies while providing remarkable insight into neuroblastoma biology. Findings provide crucial evidence in support of combining clinically approved copper chelators with immunotherapy to improve outcomes for neuroblastoma patients. Citation Format: Jourdin R. Rouaen, Daniele Mercatelli, Federica Saletta, Ensieh M. Poursani, Jayne E. Murray, Nicodemus Tedla, Federico M. Giorgi, Orazio Vittorio. Copper chelation overcomes the immunosuppressive tumor microenvironment in neuroblastoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5214.
Abstract 2285: Co-expressed gene networks in CRC CMS1 subtype
Authors: Sha'Kayla K. Nunez, Corey D. Young, Ti'ara L. Griffen, Scott Kopetz, James W. Lillard
Journal: Cancer Research (2022)

Abstract Colorectal cancer (CRC) is a heterogenous disease driven by dysregulated Wnt, Ras-Raf-MAPK, TGF-β, and PI3K-Akt signaling, which leads to an upregulation of proliferative and survival signals from the tumor microenvironment. Molecular and genomic alterations, immune signatures and gene mutations contribute to the progression of CRC. The consensus molecular subtype (CMS), an RNA-based classification system with four classes, is used to better understand CRC heterogeneity. CMS1 is the immunogenic subtype, enriched with MSI; CMS2-CMS3 has epithelial characteristics; and CMS4 is characterized by mesenchymal transitions associated gene expression. CMS1 tumors display immune infiltration, hypermethylation and microsatellite instability, and are known to have a poorer prognosis. We investigated gene signatures and associated coding mRNAs, which are co-expressed by CMS1 CRC. We acquired CRC primary tumor samples from The Cancer Genome Atlas (TCGA). CXCL13, CXCR5, IL10, PIK3R5, PIK3AP1, CCL19 and other co-expressed genes were identified to be positively correlated with CMS1. The canonical pathways and their upstream regulators of this co-expressed network were analyzed using Gene Ontology (GO). The co-expressed network associated with CMS1 was significantly and positively correlated with PI3K, WNT, ERK1 and ERK2 activation. This network was also positively correlated to right-sided tumors, microsatellite instability, chemokine-mediated signaling pathways, and immune responses. Our findings have confirmed and identified new gene signatures significantly correlated with CMS1, which in no doubt activate key pathways associated with CRC progression. Citation Format: Sha'Kayla K. Nunez, Corey D. Young, Ti'ara L. Griffen, Scott Kopetz, James W. Lillard. Co-expressed gene networks in CRC CMS1 subtype [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2285.
Abstract 226: Recurrent trisomies, variable selection and precancerous evolution in the normal gastric epithelium
Authors: Tim H. Coorens, Grace Collord, Hyungchul Jung, Yichen Wang, Sonia Zumalave, Daniel Leongamornlert, Luiza Moore, Krishnaa Mahbubani, Kourosh Saeb-Parsy, Suet Yi Leung, Michael R. Stratton
Journal: Cancer Research (2022)

Abstract Gastric cancer is the third leading cause of cancer-related mortality globally and is often associated with infectious agents such as the bacterium Helicobacter pylori. However, the background mutational landscape in normal gastric epithelium and the first genomic steps towards the formation of gastric cancer remain poorly understood. Here, we use whole-genome sequencing of microdissected gastric glands (n=271) from 30 patients, 18 of whom had gastric cancer. We show that gastric glands are clonal structures and accrue approximately 27 single base substitutions per year. While the mutational signatures in most normal glands reflect age-related mutagenesis, gastric glands sampled close to a tumor showed exposure to a mutagenic process enriched in tumors (COSMIC reference mutational signature SBS17). Phylogenetic analysis shows that acquisition of SBS17 substitutions is closely linked to overt malignant transformation. We also observe widespread trisomies of specific chromosomes, which are recurrently and independently acquired in many gastric glands of the same patient. Mutations in genes encoding epigenetic modifiers and chromatin remodelers showed evidence of positive selection and were highly enriched in some patients. This was confirmed by targeted sequencing of cancer genes in a further 1008 gastric glands. Strikingly, glands that exhibit driver mutations, a recurrent trisomy or elevated mutation loads only overlap minimally, suggesting a highly variable and patient-specific mutation and selection landscape in the normal gastric epithelium. Taken together, these results give novel insights into the preclinical evolution of gastric malignancies. Citation Format: Tim H. Coorens, Grace Collord, Hyungchul Jung, Yichen Wang, Sonia Zumalave, Daniel Leongamornlert, Luiza Moore, Krishnaa Mahbubani, Kourosh Saeb-Parsy, Suet Yi Leung, Michael R. Stratton. Recurrent trisomies, variable selection and precancerous evolution in the normal gastric epithelium [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 226.
Blockade of interleukin 10 potentiates antitumour immune function in human colorectal cancer liver metastases
Authors: Kevin M Sullivan, Xiuyun Jiang, Prajna Guha, Christopher Lausted, Jason A Carter, Cynthia Hsu, Kevin P Labadie, Karan Kohli, Heidi L Kenerson, Sara K Daniel, Xiaowei Yan, Changting Meng, Arezou Abbasi, Marina Chan, Y David Seo, James O Park, Ian Nicholas Crispe, Raymond S Yeung, Teresa S Kim, Taranjit S Gujral, Qiang Tian, Steven C Katz, Venu G Pillarisetty
Journal: Gut (2022)

Objective Programmed cell death protein 1 (PD-1) checkpoint inhibition and adoptive cellular therapy have had limited success in patients with microsatellite stable colorectal cancer liver metastases (CRLM). We sought to evaluate the effect of interleukin 10 (IL-10) blockade on endogenous T cell and chimeric antigen receptor T (CAR-T) cell antitumour function in CRLM slice cultures. Design We created organotypic slice cultures from human CRLM (n=38 patients’ tumours) and tested the antitumour effects of a neutralising antibody against IL-10 (αIL-10) both alone as treatment and in combination with exogenously administered carcinoembryonic antigen (CEA)-specific CAR-T cells. We evaluated slice cultures with single and multiplex immunohistochemistry, in situ hybridisation, single-cell RNA sequencing, reverse-phase protein arrays and time-lapse fluorescent microscopy. Results αIL-10 generated a 1.8-fold increase in T cell-mediated carcinoma cell death in human CRLM slice cultures. αIL-10 significantly increased proportions of CD8 + T cells without exhaustion transcription changes, and increased human leukocyte antigen - DR isotype (HLA-DR) expression of macrophages. The antitumour effects of αIL-10 were reversed by major histocompatibility complex class I or II (MHC-I or MHC-II) blockade, confirming the essential role of antigen presenting cells. Interrupting IL-10 signalling also rescued murine CAR-T cell proliferation and cytotoxicity from myeloid cell-mediated immunosuppression. In human CRLM slices, αIL-10 increased CEA-specific CAR-T cell activation and CAR-T cell-mediated cytotoxicity, with nearly 70% carcinoma cell apoptosis across multiple human tumours. Pretreatment with an IL-10 receptor blocking antibody also potentiated CAR-T function. Conclusion Neutralising the effects of IL-10 in human CRLM has therapeutic potential as a stand-alone treatment and to augment the function of adoptively transferred CAR-T cells.
Abstract LB008: Characterization of immune correlates of clinical activity for Nous-209, an off-the-shelf immunotherapy, with pembrolizumab for treatment of tumors characterized by microsatellite instability (MSI)
Authors: Anna Morena D'Alise, Guido Leoni, Francesca Langone, Elisa Micarelli, Gabriella Cotugno, Michael James Overman, Marwan Fakih, Dung Le, Théa Faivre, Elisa Scarselli
Journal: Cancer Research (2022)

Abstract Tumors with microsatellite instability (MSI) are caused by a deficiency in DNA mismatch repair (dMMR) system that leads to the accumulation of mutations within microsatellite regions. These types of mutations can determine a shift of the translational reading frame, generating Frame Shifted Peptides (FSPs). FSPs are tumor-specific neoantigens. Nous-209 is an off-the-shelf vaccine encoding 209 FSPs shared across dMMR/MSI tumors (Leoni et al., Cancer Res, 2020). The combination of the Nous-209 genetic polyvalent cancer vaccine and pembrolizumab has been demonstrated to be safe, highly immunogenic, with promising early signs of clinical efficacy in 12 patients with 7 durable confirmed PRs, 2 durable SDs and 3 PDs observed (Overman et al., SITC Nov 13, 2021 Poster number: 410). Here, we explore immune correlates potentially associated with the clinical outcome with the aim to address the Nous-209 contribution to the activity of pembrolizumab in this patient population. Several immunological analyses were performed on biological samples collected before and after treatment with Nous-209. Analyses were performed on paired blood and tumor biopsy samples. In periphery, vaccine immunogenicity was demonstrated by ex-vivo interferon-gamma ELISpot assay in 67% of subjects in dose level 1 (n=3), and 100% (n=7) of patients in dose level 2 with a mean of ≈1,500 IFN-γ SFCs/million PBMCs at the peak of the immune response. In 3 patients with partial response whose pre/post tumor biopsies were available, the intratumoral TCR repertoire was expanded and diversified post treatment with Nous-209. Moreover, in one out of these three patients it was possible to track vaccine-induced neoantigen specific TCR in the tumor biopsy post Nous-209 treatment. Overall, these results indicate that neoantigen specific CD8+ T cells, induced by Nous-209, expand and diversify upon treatment, and successfully traffic to and infiltrate the tumor microenvironment to exert anti-tumor activity. Citation Format: Anna Morena D'Alise, Guido Leoni, Francesca Langone, Elisa Micarelli, Gabriella Cotugno, Michael James Overman, Marwan Fakih, Dung Le, Théa Faivre, Elisa Scarselli. Characterization of immune correlates of clinical activity for Nous-209, an off-the-shelf immunotherapy, with pembrolizumab for treatment of tumors characterized by microsatellite instability (MSI) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr LB008.
Abstract 283: Reconstructed plate-based 3D screening assays that predict <i>in vivo</i> responses
Authors: Sumithra Urs, Michael Steffey, Ying Qu, Matthew Ryou, Julia Kirshner, Scott Wise
Journal: Cancer Research (2022)

Abstract In preclinical drug development, there is an urgency to create models that are predictive and relevant at the same time. To support this effort, early drug development emphasizes establishing multiple strategies that address treatment response, resistance, and combination effects, all with rapid turnover. Given the ever-increasing relevance of 3D in vitro models, Labcorp employs 3D model systems co-developed with zPREDICTA. These organ-specific models can support tumor cell growth and mimic the composition and architecture of both solid tumors and hematological malignancies. While 2D in vitro assays are preferred for screening efforts of chemical libraries, mainly because of cost, they do not recapitulate the tumor architecture and the microenvironment that is more comparable to in vivo models. This can result in differences in pharmacological potencies of standard of care (SoC) agents when compared to the 3D and in vivo models. zPREDICTA 3D assays serve as a tool for in vitro screening of tumor cell lines, which in long term cultures develop similar characteristics to in vivo disease states. Specifically, this in vitro system correlates well with breast and lung carcinoma in vivo models when matching rank order of potencies of known SoC agents. zPREDICTA tumor-specific 3D systems as well as 2D cultures and in vivo models were used to compare and contrast the activity of a set of anticancer agents. Human MDA-MB-231 cells were sensitive to paclitaxel both in 2D and in the 3D r-Breast (IC50 of 7.9nM and 6.1nM, respectively) as well as in vivo, with complete growth inhibition at the dose of 15mg/kg. Paclitaxel was cytotoxic in 2D cultures of murine 4T1 breast tumor cells (IC50=59nM), but not in the 3D reconstructed mouse breast model (r-mBreast) or in vivo, where the syngeneic 4T1 model was refractory to paclitaxel up to 10mg/kg. Similarly, NCI-H460 cells were sensitive to paclitaxel in 2D (IC50=2.3nM), while minimal response was seen in the 3D r-Lung model, and in vivo there was minimal response seen at a 20mg/kg treatment. These same cells were refractory to erlotinib in 2D, 3D, and in vivo. Additional SoC agents were profiled in these models and will also be discussed. Taken together these data demonstrate that zPREDICTA’s organ-specific 3D models mimic in vivo response with a higher fidelity than the standard 2D cultures. Additional in vitro reconstructed 3D histotype models are being developed and validated with the goal of providing faster and more predictive in vivo results. Citation Format: Sumithra Urs, Michael Steffey, Ying Qu, Matthew Ryou, Julia Kirshner, Scott Wise. Reconstructed plate-based 3D screening assays that predict in vivo responses [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 283.
Abstract 2999: Activation of NFKB is responsible for defective autophagy in cadmium-induced transformation of prostate cells
Authors: Vaibhav Shukla, Ashish Tyagi, Venkatesh Kolluru, Uttara Saran, Balaji Chandrasekaran, Murali K. Ankem, Chendil Damodaran
Journal: Cancer Research (2022)

Abstract Background: Evidence from the epidemiological, laboratory and clinical studies suggest that chronic exposure to cadmium (Cd) leads to the development and progression of prostate cancer (CaP). Previously, we reported defective autophagy due to upregulation of Placenta Specific 8 (Plac8) (a phagosome and lysosome fusion regulator) is responsible for Cd-induced transformation of normal prostate epithelial cells. This study mechanistically dissects the upstream signaling that regulates Plac8 function, which drives defective autophagy during Cd-induced transformation. Methods: To examine the molecular interaction between NFκB and Plac8, we developed NFκB/Plac8 overexpression and knockdown in normal prostate epithelial cells (RWPE-1) and cadmium transformed prostate epithelial cells lines (CTPE). In addition, we performed phenotypic, and molecular analysis including promoter-based studies, western blot, and in vivo analysis to determine the interaction between Plac8 and p65 in Cd-exposed prostate epithelial cells. Results: Our results confirmed that overexpression of Plac8 in normal prostate epithelial cells mimics Cd-transformed prostate cells (CTPE) and induces defective autophagy by regulating autophagy-related genes (ATGs) and an enhanced autophagic flux. Conversely, either NFκB or Plac8 knockdown in CTPE cells abrogates autophagy signaling. Further, Cd-exposure failed to induce defective autophagy signaling in transformed cells. Silencing p65 either by genetically or pharmacological inhibitors downregulated Plac8 expression and its survival function in CTPE cells. Hence, we investigated the interaction between NFκB and Plac8 by promoter-based analysis. We found NFκB binding site in the Plac8 promoter, and mutating NFκB binding sites in the Plac8 promoter showed a decline in luciferase activity confirming our hypothesis that NFκB activation is necessary for Plac8 activation and autophagy signaling in CTPE cells. Finally, we confirmed that silencing Plac8 significantly inhibited the tumor growth of CTPE cells compared to vehicle control. Our ongoing studies on xenotransplanted stably knockdown of p65 in CTPE cells may confirm that NFκB activation is responsible for Plac8 function and tumor growth in mice models. Conclusion: In conclusion, our results suggest that NFκB transcriptionally regulates Plac8 function, which drives pro-survival autophagy signaling in Cd-transformed prostate epithelial cells. Citation Format: Vaibhav Shukla, Ashish Tyagi, Venkatesh Kolluru, Uttara Saran, Balaji Chandrasekaran, Murali K. Ankem, Chendil Damodaran. Activation of NFKB is responsible for defective autophagy in cadmium-induced transformation of prostate cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2999.